文章摘要
孙良璋,徐正水,张丹杰,马震川,李建忠,孔冉冉,张 晋.Nampt及高糖高胰岛素微环境对人顺铂耐药肺癌细胞株增殖和转移的影响[J].,2023,(24):4620-4626
Nampt及高糖高胰岛素微环境对人顺铂耐药肺癌细胞株增殖和转移的影响
Effects of Nampt and High Glucose and High Insulin Microenvironment on Proliferation and Metastasis of Human Cisplatin-resistant Lung Cancer Cell Lines
投稿时间:2023-05-29  修订日期:2023-06-25
DOI:10.13241/j.cnki.pmb.2023.24.004
中文关键词: 烟酰胺磷酸核糖转移酶  高糖  高胰岛素  肺癌  顺铂
英文关键词: Nicotinamide phosphoribosyltransferase  High glucose  High insulin  Lung cancer  Cisplatin
基金项目:陕西省自然科学基础研究计划项目(2023-JC-QN-0840);西安市科技计划项目(2019114613YX001SF034(7))
作者单位
孙良璋 西安交通大学第二附属医院胸外科 陕西 西安 710001 
徐正水 西安交通大学第二附属医院胸外科 陕西 西安 710001 
张丹杰 西安交通大学第二附属医院胸外科 陕西 西安 710001 
马震川 西安交通大学第二附属医院胸外科 陕西 西安 710001 
李建忠 西安交通大学第二附属医院胸外科 陕西 西安 710001 
孔冉冉 西安交通大学第二附属医院胸外科 陕西 西安 710001 
张 晋 西安交通大学第二附属医院胸外科 陕西 西安 710001 
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中文摘要:
      摘要 目的:探究烟酰胺磷酸核糖转移酶(Nampt)及高糖高胰岛素(HG+HI)微环境对人顺铂耐药肺癌细胞增殖和转移的影响。方法:将人顺铂耐药细胞株A549/DDP分为6组(n=6):对照组(control)、高糖高胰岛素干预组(HH,使用添加30 mmol/L的葡萄糖和500 mU/L的胰岛素的培养基培养72 h)、分别转染sh-NC和sh-Nampt组(sh-NC和sh-Nampt,使用Lipofectamine 2000将sh-NC和sh-Nampt分别转染到细胞中,转染时间为48 h)、HH干预sh-NC和sh-Nampt组(HH+sh-NC和HH+sh-Nampt)。每组6个重复样本。qRT-PCR检测转染效率,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell检测细胞迁移和侵袭,qRT-PCR检测Nampt mRNA,Western blot检测Nampt、Bcl-2、Bax、MMP-2、MMP-9、p-PI3K、PI3K、p-AKT和AKT蛋白表达。结果:与对照组和sh-NC组比较,sh-Nampt组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量降低,而细胞凋亡率升高,Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平降低,Bax蛋白表达水平升高(P<0.005)。与对照组和sh-NC组比较,HH组和HH+sh-NC组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量升高,Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平升高,Bax蛋白表达水平降低(P<0.005)。与HH组和HH+sh-NC组比较,HH+sh-Nampt组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量降低,细胞凋亡率升高,Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平降低,Bax蛋白表达水平升高(P<0.005)。结论:高糖高胰岛素微环境可能通过上调Nampt/PI3K/AKT信号通路诱导人顺铂耐药肺癌细胞的增殖和转移。
英文摘要:
      ABSTRACT Objective: To reveal the effects of nicotinamide phosphoribosyltransferase (Nampt) and high glucose and high insulin (HG+HI) microenvironment on proliferation and metastasis of human cisplatin-resistant lung cancer cell lines. Methods: Human cisplatin resistant cell line A549/DDP was divided into 6 groups (n=6): control group, high glucose and high insulin intervention group (HH, cultured with 30 mmol/L glucose and 500 mU/L insulin for 72 h), sh-NC and sh-Nampt transfected groups (Lipofectamine 2000 was used to transfect sh-NC and sh-Nampt into cells respectively, the transfection time was 48 h), HH intervention sh-NC and sh-Nampt groups (HH+sh-NC and HH+sh-Nampt). Each group had 6 replicates. The transfection efficiency was detected by qRT-PCR. Cell proliferation was detected by MTT method, apoptosis was detected by flow cytometry, cell migration and invasion were detected by Transwell, Nampt mRNA was detected by qRT-PCR, and protein expression of Nampt, Bcl-2, Bax, MMP-2, MMP-9, p-PI3K, PI3K, p-AKT and AKT were detected by Western blot. Results: Compared with control group and sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells decreased, the rate of apoptosis increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein decreased, and the expression level of Bax protein increased in sh-Nampt group (P<0.05). Compared with control group and sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein increased, and the expression level of Bax protein decreased in HH group and HH+sh-NC group(P<0.05). Compared with HH group and HH+sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells decreased, the rate of apoptosis increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein decreased, and the expression level of Bax protein increased in HH+sh-Nampt group(P<0.05). Conclusion: High glucose and high insulin microenvironment may induce proliferation and metastasis of human cisplatin-resistant lung cancer cells by up-regulating Nampt/PI3K/AKT signaling pathway.
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